Isolation of chloramphenicol acetyltransferase from soil bacteria for sterility testing of products containing chloramphenicol.

ABSTRACT

For large scale isolation of chloramphenicol acetyltransferase (CAT), five soil bacteria (Alkaligens faecalis cc; Escherichia coli c-18; Escherichia coli c-22; Escherichia coli c-24 and Klebsiella pneumoniae c-38) resistant to chloramphenicol (Cm) were tested with surface active agents such as sodium dodecyl sulphate (SDS), triton x-100, tween-80 and sodium deoxycholate (SDC). CAT leakage was satisfactory with SDS and SDC. Since SDS forms a covalent complex with certain proteins, extraction of CAT was performed with SDC in the present study. The enzyme was partially purified by a gel filtration procedure. The stability test showed that the CAT enzymes isolated from E. coli c-22 and K. pneumoniae c-38, were fairly stable even at room temperature. The CAT isolated in this way may be useful for sterility testing of Cm containing products.